ARTICLE IN PRESS

WAT E R R E S E A R C H 40 (2006) 2007 – 2016

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Treatment of olive oil mill wastewater by combined process

electro-Fenton reaction and anaerobic digestion

Sonia Khoufi, Fathi Aloui, Sami Sayadi

Laboratoire des Bioprocédés, Centre de Biotechnologie de Sfax; B. P. ‘‘K’’, 3038 Sfax, Tunisia

art i cle info A B S T R A C T

Article history: In this work, we investigated an integrated technology for the treatment of the recalcitrant
Received 23 October 2005 contaminants of olive mill wastewaters (OMW), allowing water recovery and reuse for
Received in revised form agricultural purposes. The method involves an electrochemical pre-treatment step of the
19 March 2006 wastewater using the electro-Fenton reaction followed by an anaerobic bio-treatment. The
Accepted 26 March 2006 electro-Fenton process removed 65.8% of the total polyphenolic compounds and subse-
quently decreased the OMW toxicity from 100% to 66.9%, which resulted in improving the
Keywords: performance of the anaerobic digestion. A continuous lab-scale methanogenic reactor was
Olive mill wastewater operated at a loading rate of 10 g chemical oxygen demand (COD) l
1 d
1 without any
electro-Fenton apparent toxicity. Furthermore, in the combined process, a high overall reduction in COD,
Anaerobic digestion suspended solids, polyphenols and lipid content was achieved by the two successive
Polyphenols stages. This result opens promising perspectives since its conception as a fast and cheap
pre-treatment prior to conventional anaerobic post-treatment. The use of electro-
Abbreviations:
coagulation as post-treatment technology completely detoxified the anaerobic effluent
OMW: olive mill wastewaters
and removed its toxic compounds.
COD: chemical oxygen demand
& 2006 Elsevier Ltd. All rights reserved.
BOD5: biological oxygen demand
TSS: total suspended solids
LMM: low molecular mass
AF: anaerobic filter
VFA: volatile fatty acids

1. Introduction as well as their acidity and high concentration of potassium,

Treatment and disposal of olive mill wastewater (OMW)
represents one of the main problems for olive oil producing
countries of the Mediterranean area. Tunisia is one of the
largest olive oil producers in the world with an average
annual production of 450,000 tons. This results in a by-
product of 600,000 m3 OMW. These liquid residues are
100–150 times more heavily loaded with pollutants than
ordinary domestic wastewater (Sabbah et al., 2004). The high
polluting activity of OMW is linked with their high content of
organic molecules, especially polyphenolic mixtures
(4–10 g l
1) with different molecular weights (Hamdi, 1992),
magnesium and phosphate salts (Arienzo and Capasso, 2000).
Besides aromatic compounds, OMW contain other organic
molecules including nitrogen compounds, sugars, organic
acids, and pectins (Della Greca et al., 2000), that increase their
organic load (chemical oxygen demand (COD) ¼ 80–200 g l
1;
biological oxygen demand (BOD5) ¼ 50–100 g l
1). Further-
more, the physico-chemical characteristics of OMW are rather
variable, depending on climatic conditions, olive cultivars,
degree of fruit maturation, storage time, and extraction
procedure.
Many pollution disposal methods, such as concentration,
evaporation, incineration, ultrafiltration/reverse osmosis,

Corresponding author. Tel./fax: +00 216 74 440 452.

E-mail address: sami.sayadi@cbs.rnrt.tn (S. Sayadi).
0043-1354/$ - see front matter & 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.watres.2006.03.023
 ARTICLE IN PRESS
2008 WAT E R R E S E A R C H
lime precipitation, aerobic treatment, lagooning, codigestion,
etc were tested on OMW, none of them led to industrial
applications. However, anaerobic digestion seems to have
some clear advantages that would make it the process of
choice (Borja et al., 1992). Indeed, this treatment process
produces energy (methane) and a digested effluent with a
significant reduction of the organic load (Marques, 2001).
However, many problems concerning the high toxicity and
inhibition of biodegradation of these effluents were encoun-
tered during anaerobic treatments, because some bacteria,
such as methanogens, were particularly sensitive to the
organic contaminants present (Andreoni et al., 1984). The
phenolic compounds severely limit the possibility of using
anaerobic digestion (Sayadi et al., 2000). Therefore, the
elimination of phenolic compounds from OMW was consid-
ered as an important objective in order to reduce its toxicity
and to permit the occurrence of microbial fermentation. For
this, research turned to a more promising alternative, namely
the physico-chemical pre-treatment to remove the toxic
compounds of OMW (Beccari et al., 1999).
In recent years, there has been increasing interest in the
use of electrochemical technologies for the treatment of
wastewaters. This technique was found to be successful in
removing pollutants in various industrial wastewaters (Lin
and Chang, 2000; Ciardelli and Ranieri, 2001; Lai and Lin,
2004). In two recent investigations, Inan et al., (2004) and
Adhoum and Moncer (2004), an electrochemical method was
used for decreasing the organic matter in OMW. Both
investigators found efficient removals of COD, colouration
and polyphenols content by electrolysis process using alumi-
nium and iron electrodes. However, a relatively new chemical
oxidation method that has not received much attention for
OMW or other industrial wastewater treatment is the electro-
Fenton method (Lin and Chen, 1997). This method represents
a combination of the electrochemical process and the Fenton
oxidation. It is based on the fact that hydrogen peroxide
(H2O2) can be used as an oxidant in advanced oxidation
processes to decompose refractory or toxic wastewaters
(Kusvuran et al., 2004). As indicated in reaction (1), when
the ferrous ion reacts with H2O2 it will generate strong
oxidant hydroxyl radicals (OH  ).
Fe2þ þ H2 O2 ! Fe3þ þ OH þ OH
: (1)
This Fe2+/H2O2 system, often referred to as Fenton’s reagent
(Fenton’s), has dual functions of OH radical peroxidation as
well as ferrous/ferric coagulation.
During this process, the non-biodegradable organics and
toxic pollutants present in the wastewaters such as poly-
phenols are usually destroyed by direct or indirect anodic
oxidation via the production of oxidants such as hydroxyl
radicals and complex coagulants that promote the floccula-
tion of the matter (Israilides et al., 1997; Panizza et al., 2000;
Chen et al., 2002).
This paper will attempt to apply the electro-Fenton process
to reduce the organic load and the toxicity of OMW in order to
improve the anaerobic digestion in terms of biomethane
yield. Electro-coagulation was assayed as a post-treatment for
complete detoxification and colour removal allowing water
recovery and reuse for agricultural purposes.
40 (2006) 2007– 2016
2. Materials and methods
2.1. OMW characterisation
Fresh OMW was obtained from an olive oil continuous
processing plant located in Sfax (southern Tunisia). The
OMW was characterised by high total suspended solids
(TSS) content, COD concentration up to 100 g l
1 and poly-
phenols up to 12 g l
1. Raw OMW was pre-decanted in a 120 l
decanter before being treated by electro-Fenton in order to
remove suspended solids (Fig. 1).
To confirm the role of electro-Fenton in polymerising and
removing the highly polymerised phenolic fraction, experi-
ments were carried out with a low-molecular-mass (LMM)
polyphenolic fraction (o2 kDa) obtained by the ultrafiltration
of crude OMW using a polysulphone 2 kDa cut-off membrane.
The purpose of this ultrafiltration was to study the effect of
electro-Fenton reaction on the toxic fraction of OMW which is
composed of LMM phenolics such as simple phenolics
(hydroxytyrosol, tyrosol, p-OH benzoic acid, p-OH phenyl
acetic acid, vannilic acid, caffeic acid, coumaric acid, vanni-
line, ferilic acid, catechol, methylcatechol), tannins, antocya-
nins, catechin (Sayadi et al., 2000). The C18-HPLC
chromatogram of this OMW phenolic fraction is presented
in Fig. 2.
2.2. Electro-Fenton and electro-coagulation treatment
Preliminary experiments were carried out in a 0.25 l
1 glass
reactor for the electro-Fenton of OMW fraction. The aqueous
solution of reactants was homogenised by magnetic agitation
to avoid concentration gradients. The electro-Fenton reactor
was formed by one pair of anodic and cathodic electrodes
(cast iron plates) which were positioned approximately 1.5 cm
apart from each other and were dipped in the effluent. The
total effective surface area of electrodes was 0.2 dm2. The
current input was supplied by a convergy power supply. In
each run, approximately 0.2 l of OMW fraction was placed in
the electrolytic cell. The pH of the solution was adjusted to 4.
H2O2 was added to the electrolytic cell before the electrical
current was turned on. A batch study was conducted to
optimise parameters like H2O2 concentration and current
density governing the electro-Fenton process. These para-
meters were examined in the range of 0–1.5 g l
1 and
1.25–10 A dm
2, respectively. The optimum H2O2 concentra-
tion and current density were found to be 1 g l
1 and
7.5 A dm
2, respectively. At these conditions, maximum
removal of monomer concentration, COD and colour were
attained. For this reason, these conditions were chosen as the
optimised parameters and were subsequently used for
preparing the pre-treated OMW for the biomethanisation.
Experiments of electro-Fenton of crude OMW were conducted
in a 5 l glass reactor using iron electrodes having an effective
surface area of 150 dm2 (Fig. 1). In each run, 3 l of crude OMW
were treated and operated in batch mode.
Electro-coagulation of anaerobic effluent was carried out in
the same reactor as for the electro-Fenton of crude OMW
without stirring. This electrolysis process lasted 2 h at
1.8 A dm
2 and without adjustment of pH.
 ARTICLE IN PRESS
WAT E R R E S E A R C H 4 0 (200 6) 200 7 – 201 6 2009

Fig. 1 – Schematic representation of OMW treatment process.

Fig. 2 – C18-HPLC chromatogram of phenolic compounds present in OMW fraction. 1: Hydroxytyrosol; 2: 3, 4 dihydroxyphenyl
acetic acid, 3: tyrosol; 4: p-OH benzoı̈c acid; 5: p-OH phenylacetic acid; 6: vanillic acid; 7: caffeic acid; 8: coumaric acid; 9:
vanillin; 10: ferulic acid.

After electro-Fenton reaction and electro-coagulation, trea-
ted effluents were placed in decanter tanks (Fig. 1) to
eliminate sludge formed during electrolysis. During the
experiments, samples were withdrawn and immediately
analysed for water quality measurements.
2.3. Anaerobic biotreatment and biogas analysis
Two anaerobic filters (AFs) were used in this study. These
reactors were made of a glass column having a working
volume of 3 l. The inner tubes were enclosed in a jacket
through which hot water was circulated to maintain the
temperature of the filter at 37 1C. These AFs were packed with
polyurethane foam cubes 2 cm  2 cm  1 cm (Filtren T45,
from Recticel, Wetteren, Belgium) as support and inoculated
with an 8-year-old digester operated with pre-treated OMW.
The influent was fed in six times into the reactor using a
pump connected to a programmer. For monitoring the volatile
fatty acids (VFA) inside the reactor, three sampling points
were made in the AF. Level (A) was at the bottom of the
reactor. Level (B) corresponded to the middle and level (C) was
at the top of the reactor.
 ARTICLE IN PRESS
2010 WAT E R R E S E A R C H
The gas flow rates of the AFs were measured by liquid
displacement. Gas samples were taken with a syringe from
the tank of biogas. CH4, CO2 and N2 were measured using a
gas chromatograph GC11 (Delsi instruments) equipped with a
Haye SepQ 60/80 (SUPELCO) column (maintained at 60 1C), a
thermal conductivity detector (current intensity of 160 mA)
and a servotrace integrator (SEFRAM). Helium was used as a
carrier gas at a pressure of 1.3 bar.
2.4. Analytical methods
As the presence of residual H2O2 introduces a positive error in
COD determination (Kang et al., 1999), in electrolysis
experiments with H2O2, the pH of the samples was raised to
above 10 with NaOH 6 N prior to analysis. The value of
COD was estimated using the method described by Knechtel
(1978) and fading colour was monitored by measuring the
absorbance at 395 nm, the length of the maximum absor-
bance, using a spectrophotometer (ANTHLIE ADVANCED
5 SECOMAM). Samples were centrifuged for 20 min at
4000 t/min and diluted appropriately before each COD deter-
mination.
BOD5 was determined by the manometric method with a
respirometer (BSB-Controller Model 620T (WTW)).
Concentration of ortho-diphenols was determined by the
colorimetric reaction with Folin–Ciocalteau reagent. An
aliquot of the OMW aqueous methanol extract was mixed
with 2 ml of Folin–Ciocalteau reagent (Fluka, Switzerland). A
sodium hydroxide solution (6% v/v) was added, and the
mixture was shaken. The blue colour formed was measured
at 727 nm. The ortho-diphenol concentration of OMW sam-
ples, as determined by the Folin–Ciocalteau method, (Folin
and Ciocalteau, 1927) were reported as caffeic acid equiva-
lents by reference to a standard curve. As about total
polyphenols, they were quantified using the method de-
scribed by Sayadi et al. (2000).
Concentration of aromatic compounds was determined by
high-performance liquid chromatography (HPLC) using a
Shimadzu 10AVP chromatograph equipped with a Shimadzu
10AVP UV detector. Separation was made by a column (Shim-
pack CLC-ODS (M) 250 mm  4.6 mm) washed with acetoni-
trile/water (70/30) before and after analysis. A mixture of 50%
acetonitrile in 50% water was chosen as optimal mobile
phase. Data were analysed by class VP Shimadzu software.
A Progel TSK-G 2000-SW Supelco column (300 mm 
7.8 mm) was used with the same Shimadzu apparatus to
analyse molecular-mass distribution of the OMW polyphe-
nols. The elution was carried out using a phosphate buffer of
pH 6.8 and 0.6 ml min
1 flow rate. The wavelength of the
detector was adjusted to 280 nm.
The standard method of Soxhlet solid/liquid (organic solids
of OMW/hexane) was utilised for the dosage of lipids.
VFAs (acetate, propionate, butyrate, isobutyrate and vale-
rate) were measured by HPLC using the method described by
Mechichi and Sayadi (2005).
The microtoxicity test consists of the inhibition of the
bioluminescence of Vibrio fischeri LCK480 using the LUMIStox
system (Dr Lange GmbH, Düsseldorf, Germany) and was
carried out according to ISO 11348-2 (1998). Percentage
inhibition of the bioluminescence was achieved by mixing
40 (2006) 2007– 2016
0.5 ml of OMW and 0.5 ml luminescent bacterial suspension.
After a 15 min exposure at 15 1C, the decrease in light
emission was measured. The toxicity of the OMW is
expressed as the percent of the inhibition of bioluminescence
(%IB) relative to a non-contaminated reference. A positive
control (7.5% NaCl) was included for each test.
Phytotoxicity test was estimated by the determination of
the germination index (GI) according to Wong et al. (2001)
using Lycopersicon esculentum (tomato) seeds.
3. Results and discussion
3.1. Electro-Fenton treatment
3.1.1. Electro-Fenton treatment of OMW fraction
Preliminary tests were conducted to study the effect of
electro-Fenton reaction on the pollutant characteristics of
the LMM phenolic fraction. Experiments were realised with
1 g l
1 H2O2 added and a current density of 7.5 A dm
2 (see
materials and methods section). The initial pH of the OMW
fraction (4.8) was decreased to pH 4 in order to allow the
Fenton’s peroxidation. Fig. 3 shows the evolution of pH, COD
removal, colouration, monomers removal, hydroxytyrosol
concentration and toxicity using V. fischeri based on LUMISTox
system during the electro-Fenton treatment. The pH in-
creased from 4 to 9 (Fig. 3a). The final pH differed in function
of the quantity of the H2O2 added (data not shown) and the
duration of the treatment. For 1 g l
1 of H2O2 and a 6 h
treatment, the final pH obtained (pH 10.5) was not favourable
for anaerobic post-treatment. The COD removal was 26% (Fig.
3b) at the steady state (after 4 h) but could be higher for a
prolonged treatment period. The colour intensity of the
effluent fraction monitored by measuring absorbance at
395 nm (Fig. 3c) doubled after 30 min of reaction, probably
due to the polymerisation of monoaromatic compounds.
Then it decreased to 75% of the initial colour after 4 h. This
can be explained by the coagulation of the highly polymerised
polyphenolic compounds.
Chromatography techniques confirmed the removal of the
most phenolics of LMM (Fig. 3d) which resulted in decreasing
the toxicity from 100% to 67% after 30 min (TF1) and to 28%
after 4 h of incubation (TF2) (Fig. 3f). Indeed, approximately
90% of the mono-aromatic compounds were removed after
4 h of incubation. As an example, the concentration of
hydroxytyrosol, the major ortho-diphenol, decreased rapidly
in the OMW fraction (Fig. 3e); a removal of 98% was achieved
at the end of treatment.
To test whether the dark colour was caused by
polymerisation of the OMW fraction, the molecular-mass
distribution of the reaction mixtures was measured by fast
SE-HPLC and compared with the control. A polyphenol
fraction with high hydrodynamic volumes was formed after
OMW oxidation by electro-Fenton oxidation, suggesting that
polymerisation had taken place (Fig. 4). After this polymer-
isation step which was related to the colour increase, a
coagulation of the highly condensed polymers followed by
rapid sedimentation occurred. This resulted in the decolour-
isation of the OMW fraction.
 ARTICLE IN PRESS
WAT E R R E S E A R C H 4 0 (200 6) 200 7 – 201 6 2011

10 100

9 80

Maximum removal (%)

8
60
pH

7
40
6
20
5

4 0
0 1 2 3 4 1 2 3 4 5 6 7 8 9 10
(a) Time (h) (d) Monomers

30 1000

concentration (mg l-1)

800
COD removal (%)

Hydroxytyrosol
20
600

400
10
200

0 0
0 1 2 3 4 0 1 2 3 4
(b) Time (h) (e) Time (h)

250 100
Residual coloration (%)

Relative inhibition (%)

200 80

150 60

100 40

50 20

0 0
0 1 2 3 4 F
TF1
(c) Time (h) (f) TF2

Fig. 3 – Variations of pH (a), COD removal (b), residual colour (c), monomer removal (d) (1: hydroxytyrosol; 2: 3, 4 di-OH
phenylacetic acid, 3: tyrosol; 4: p-OH benzoı̈c acid; 5: p-OH phenylacetic acid; 6: vanillic acid; 7: caffeic acid; 8: coumaric acid;
9: vanillin; 10: ferulic acid), hydroxytyrosol concentration (e) and the relative toxicity of OMW fraction (F) and the reactants
collected during the Fenton process at 30 min (TF1) and at 4 h (TF2) of treatment (f).

3.1.2. Electro-Fenton treatment of crude OMW
Table 1 shows the characteristics of OMW before and after
electro-Fenton treatment. After decantation, the TSS of raw
OMW decreased from 59 to 12 g l
1. The residual TSS in crude
OMW was unsettlable suspended matter, which presents a
major difficulty in the treatment and handling of OMW.
During electro-Fenton treatment, pH increased from 4 to 7.6,
which may be attributed to the smaller production of H+ than
OH
as was explained by Israilides et al. (1997) and the
reduction in phenol concentration. Indeed, phenols are acids
in liquids, and their removal from a solution reduces its
acidity. The pH value of electro-Fenton-treated OMW can be
considered favourable for anaerobic bio-treatment.
Biodegradability is determined by measuring the ratio
between COD and BOD5, whose value must be in the range
of 2–2.5. After electro-Fenton, the COD of crude OMW drops to
approximately 68% of the initial value. This result points out
the ability of the electrolysis process to eliminate soluble
compounds present in OMW. BOD5 values decreased from
19.25 to 15.5 g l
1 before and after treatment, respectively.
Thus, COD/BOD5 ratio decreased from 5.84 before to 2.26 after.
It appears that a significant proportion of the non-biodegrad-
able matter present in OMW was removed by electro-Fenton.
Degradation and mineralisation of phenolic compounds can
occur during Fenton reaction. Kavitha and Palanivelu (2004)
reported that in Fenton process, biodegradable aliphatic
 ARTICLE IN PRESS
2012 WAT E R R E S E A R C H 40 (2006) 2007– 2016

Fig. 4 – Molecular-mass distribution of phenolics from the untreated ultra-filtrated OMW phenolic fractions (___) and treated
fraction ( ). Arrows indicate standard elution times: from left to right: 1: blue dextran (MW ¼ 2000 kDa), 2: lysozym
(MW ¼ 15 kDa), 3: syringic acid (MW ¼ 198 Da).

Table 1 – Compositions of olive mill wastewaters before and after the treatment with the electro-Fenton process (S1, see
Fig. 1) then with anaerobic bio-treatment (S2, see Fig. 1) and finally by electro-coagulation (S3, see Fig. 1)

Parameter Crude OMW S1 S2 S3

pH 5.44 7.60 7.80 9.20

Color (absorbance 395 nm) 73.00 16.10 13.16 1.19
UV absorbance 280 nm 167.30 54.20 49.60 4.60
BOD5 (g l
1) 19.25 15.50 — —
COD (g l
1) 112.50 36.00 8.30 2.50
COD/BOD5 5.84 2.26 — —
Total solids (%) 12.20 3.90 1.95 0.97
Total volatiles (%) 10.90 2.10 1.50 0.65
Total suspended solids (g l
1) 59.00 2.70 1.80 0.53
Volatiles suspended solids (g l
1) 55.17 2.30 1.65 0.32
Ortho-diphenols (mg l
1) 6025.50 1536.70 861.80 28.57
Total polyphenols (g l
1) 11.75 4.20 1.20 Not detected
Residual oils (g l
1) 12.00 1.30 Not detected Not detected

compounds such as acetic acid and oxalic acid were identified
as the major products during the degradation of synthetic
phenol. However, transformation of phenolic polymers to
simple phenolic compounds was not demonstrated.
Chromatography analysis (data not shown) confirmed the
removal of most LMM phenolics. Besides, the concentration
of ortho-diphenols, monitored by Folin–Ciocalteau method,
was significantly reduced during the electro-Fenton process.
Removal efficiency was about 65.8% for total polyphenols and
74.5% for ortho-diphenols.
Crude OMW was highly coloured due to its high content of
polyaromatic compounds. In the beginning of the electrolysis
treatment, the colour intensity of the effluent increased
(data not shown) as a result of phenolic compounds
polymerisation. However, colour intensity decreased to 78%
of the initial colour at the end of treatment.
During the electrolysis treatment, a part of the solute and
particle matter present in OMW turned out to be a suspended
solid that could reach 40 g l
1 at the end of the electrolysis
reaction. These TSS were rapidly eliminated by simple
sedimentation. After decantation, the obtained effluent has a
weak quantity of TSS (2 g l
1) in comparison with the decanted
crude OMW (12 g l
1). The formation of suspended particles
was caused presumably by electro-coagulation process. The
polymers were precipitated with iron which was continuously
dissolved into the wastewater from the cast iron anodes, as
governed by the Faraday’s law (Pletcher and Walsh, 1990).
This result confirms the hypothesis that the electro-Fenton
reaction would have a strong ability to eliminate polyphenols
from OMW. Furthermore, as can be seen in the Table 1, the
concentration of lipids was decreased by 89.2%. The pH, COD,
colouration, polyphenols and lipids removal were consis-
tently very good. Indeed, the effluent quality of the
pre-treated OMW by electro-Fenton process (S1, see Fig. 1)
was rather excellent (Table 1). It could be directly fed as
influent to anaerobic reactor.
3.2. Anaerobic bio-treatments
3.2.1. Anaerobic digestion of non pre-treated OMW
The anaerobic treatment of non pre-treated OMW was
performed in a 3-l AF reactor. The yield of methanisation of
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WAT E R R E S E A R C H 4 0 (200 6) 200 7 – 201 6 2013

this untreated diluted OMW was higher than 0.3 l CH4 g
1 COD (mean value). The hydraulic retention time (HRT) varied
introduced at low loading rates. However, since the 26th day, between 17.7 and 3.5 days. In general, the percentage of
when the loading rate reached a mean of 4 g l
1 d
1 of COD, a COD removal decreased with increased loading rate during
decrease in the biogas production and yield was observed the fermentation of OMW in the AF. The percentage of COD
(Fig. 5A). This toxicity was accompanied by a pH decrease in removal decreased from 88.8% to 68% when the organic
the three levels of the reactor and an accumulation of the VFA loading rate increased from 2 to 10 g COD l
1 d
1 (Fig. 7). The
(Fig. 6). This test of the anaerobic digestion of untreated OMW evolutions of the loading rate, biogas productivity and
by an 8-year OMW-acclimated consortium will serve as a methane yield are presented in Fig. 5B.
control for comparing the efficiency of the electro-Fenton pre- At the higher loading rates (9–10 g COD l
1 d
1), the yields
treatment in the detoxification of this effluent. obtained were approximately 0.3 l CH4 g
1 COD introduced.
The volume of biogas reached 12 l d
1 (4-fold of the volume of
3.2.2. Anaerobic digestion of electro-Fenton pre-treated OMW the digester). The higher values of yields (0.32–0.34 l CH4 g
1
The AF was loaded with undiluted pre-treated OMW at a COD introduced) were obtained for loading rates lower than
starting loading rate of 2 g COD l
1 d
1. The reactor was 8 g COD l
1 d
1. In addition, Fig. 5Ba and Bc show that the
operated at influent OMW concentration of 35.5 g COD l
1 methane yield increased with the increase of the loading rate

10
10
Ba

Loading rate (g COD l-1 d-1)

Loading rate (g COD l-1 d-1)

Aa
8
8

6 6

4 4

2 2

0 0
0 20 40 60 0 20 40 60 80 100
Time (days) Time (days)

5 5
Ab Bb
4 4
Biogas (l l-1)

3 3
Biogas (l l-1)

2 2

1 1

0 0
0 20 40 60 0 20 40 60 80 100
Time (days) Time (days)

0.4 0.4
Ac Bc

0.3
Methane yield (Y)

0.3
Methane yield (Y)

0.2 0.2

0.1 0.1

0 0
0 20 40 60 0 20 40 60 80 100
Time (days) Time (days)

Fig. 5 – Evolution of the loading rate (g COD l
1 reactor d
1) (a), biogas production (l l
1) (b) and methane yield (l CH4 g
1 COD
introduced) (c) during anaerobic digestion of crude OMW (A) and electro-Fenton pre-treated OMW (B).
 ARTICLE IN PRESS
2014 WAT E R R E S E A R C H 40 (2006) 2007– 2016

10 10

9 A B C 9 A B C

8 8
pH

pH
7 7

6
6
5
5
4
4 0 20 40 60 80 100
0 20 40 60
(a) Time (days)
(a) Time (days)
10
15
A B C 8 A B C
12

VFA gl-1
6
9
VFA gl-1

4
6
2
3
0
0 20 40 60 80 100
0
0 20 40 60 (b) Time (days)
(b) Time (days) Fig. 8 – Evolution of pH (a) and concentration of VFA (b) in the
anaerobic filter during the methanisation of electro-Fenton-
Fig. 6 – Evolution of pH (a) and concentration of VFA (b) in the
pre-treated OMW (A: bottom of the reactor, B: middle of the
anaerobic filter during the methanisation of crude OMW (A:
reactor, C: top of the reactor).
bottom of the reactor, B: middle of the reactor, C: top of the
reactor).

the three levels of the AF (Fig. 8). Level A is at the bottom of

50 the reactor, level B corresponds to the middle and level C is in
COD inf COD eff the top of the reactor.
40
The pH at these three levels of the reactor was higher than
COD g l-1

30
20
10
0 that electro-Fenton of OMW resulted in decreasing the toxic
0 20 40 60 80
Time (days)
Fig. 7 – Evolution of the COD of influent and effluent of the
anaerobic filter alimented by electro-Fenton-treated OMW.
up to the theoretical steady yield value, reported to be around
0.35 l CH4 g
1 COD introduced. This observation can be
considered a solid proof for the ability of the anaerobic
biomass to degrade most organic matter present in the
electro-Fenton-pre-treated OMW. Besides, it may confirm
the gradual increase of the methanogenic activity.
The biomethanisation process was found to be stable
during 3 months of operation. No toxicity phenomenon was
observed. VFAs have long been recognised as the most
important intermediates in the anaerobic process and were
proposed as a control parameter (Ahring et al., 1995; Mechichi
and Sayadi, 2005). Therefore, VFA and pH were analysed in
7.0 for all the loading rates applied. The VFA concentrations
were low even at the higher loading rates. Knowing that
untreated OMW causes inhibition of methanisation at a
loading rate of 2–4 g COD l
1 d
1 (Kang and Chang, 1997;
Hamdi, 1991; Rozzi et al., 1989, this work), it can be concluded
100 effect of this wastewater on anaerobic digestion. Moreover,
this experiment was stopped at a loading rate of
10 g COD l
1 d
1 while the biological process did not show
any apparent toxicity. These results also suggest that
anaerobic digestion can be a practical alternative for the
treatment of OMW.
3.2.3. Characterisation of the anaerobic effluent
The anaerobic effluent (S2) was characterised with common
parameters (pH, COD, colouration, TSS, ortho-diphenols).
Main results were plotted in Table 1. Results showed that
the colouration and the residual COD (hardly biodegradable
compounds) of S2 remained relatively high. The phytotoxicity
test of OMW samples were carried out using the germination
index (GI) of L. esculentum (tomato). Results showed that
electro-Fenton treatment increased the GI percentage of
L. esculentum from 4.4% (for crude OMW) to 30% while
the anaerobic effluent led to an increase of the GI to
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Table 2 – Inhibition of Vibrio fisheri luminescence (IB) after
exposure with different OMW samples during 15 min
and the GI percentage of Lycopersicon esculentum
OMW sample IB (%) GI (%)
Untreated OMW 100 4.4
Electro-Fenton-treated OMW (S1) 66.9 30
Electro-Fenton anaerobic-treated OMW (S2) 45.2 121
Electro-coagulated S2 (S3) — 140
121% compared to 100% for the control (Table 2). Indeed,
approximately 43.9% of the ortho-diphenols were removed
after anaerobic bio-treatment.
As shown in Table 2, untreated OMW exercised 100%
inhibition on V. fischeri. It was reduced to 66.9% after pre-
treatment by electro-Fenton and to 45.2% in the anaerobic
effluent. Microtoxicity of S2 remained high due to the residual
VFA. Yet, the characteristics of anaerobic effluent do not
comply with legal requirements. To overcome this problem, a
tertiary treatment step was necessary if we want to reach the
Tunisian standard requirements. For this purpose, experi-
ments of electro-coagulation of S2 were carried out using the
same electro-Fenton reactor, in order to remove the residual
polyphenols, COD and colour.
3.3. Improvement of the quality of the effluent using
electro-coagulation
The purpose of this part of study was directed to treat the
anaerobic effluent (S2) by electro-coagulation process. During
this process, when direct current passed though the Fe
anodes, Fe2+ and Fe3+ correspondingly dissolved and com-
bined with hydroxyl ions in the water. They formed metal
hydroxyls ions, which are partly soluble in water under
definite pH values and play the role of coagulant.
The electro-coagulation step was performed at a current
density of 1.8 A dm
2 and without adjustment of pH. The
determination of the physico-chemical parameters of the
electro-coagulated anaerobic effluent (S3) showed that the
electrolysis process was able to remove 70.55% of TSS, 91% of
the colour and 70% of the residual COD (Table 1). Moreover,
the analysis of ortho-diphenols showed a removal efficiency
of 97% while polyphenolic compounds were not detected.
Hence, the final effluent (S3) was free of toxic compounds as
can be seen in Table 1. Furthermore, the phytotoxicity of S3
was significantly reduced by the application of electro-
coagulation, reaching 140% germination index (Table 2). As
conclusion, the proposed process of OMW treatment reduces
significantly its biotoxicity. For this, OMW can be used as
fertiliser.
4. Conclusion
The electro-Fenton method applied on raw OMW as pre-
treatment resulted in removing a large amount of recalcitrant
polyphenolic compounds as well as in decreasing toxicity.
4 0 (200 6) 200 7 – 201 6 2015
The anaerobic process applied as post-treatment reached a
loading rate of 10 g COD l
1 d
1 without any apparent toxicity.
Finally, electro-coagulation of the anaerobic digestion effluent
could be used as polishing step for improving the quality of
the treated water for potential reuse.
Acknowledgements
This research was supported by C.I.U.F. (Belgium), EEC
Contract no. ICA3-CT-2002-00034 and Contract Programmes
(MRSTDC, Tunisia).
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